22 August 2006
Introduction to Proteins II
Dr. Kandice Williams, Ph.D.



Primary Structure


  • Each protein has a unique amino acid sequence that defines its primary structure
  • Linking of amino acids in a linear polypeptide chain
  • α-carboxyl group of one amino acid is covalently linked to the α-amino group of another via peptide bond
    • The amino terminal end is the beginning of the polypeptide chain; carboxyl terminal end is the end of the polypeptide chain
    • Two amino acids linked together is called a dipeptide
    • Chain of more than two amino acids is called a polypeptide chain
    • All polypeptide chains are polar
  • Requires input of free energy (+21 kJ/mol) and loss of a water molecule to created a peptide bond
  • Requires +356 kJ/mol to break a peptide bond – very stable bond!

Molecular Weight

  • Molecular Weight (MW) is measured in Daltons which is equivalent to the atomic mass
  • Average MW of one amino acid is 110 Daltons
  • Most human proteins contain 50-2000 amino acid residues
    • MW would range between 5,500-220,000 Daltons

Polypeptide Chain

  • Polypeptide chain contains a peptide bond backbone and distinctive –R group side chains
    • Backbone always contains a carbonyl group, a good hydrogen bond receptor, an amine group (except for proline), and a good hydrogen bond donor
    • Backbone can interact with each other and with side chain functional groups
  • Disulfide bonds result in covalent cross-linking between and within proteins
    • -SH group forms disulfide bonds via oxidation
    • Two cysteine residues covalently bonded forms one cystine

Peptide Bonds

  • Peptide bond is essentially planar because of rigid double-bond-like characteristics
  • Peptide bond is uncharged, but is rich in hydrogen bond potential
  • For each pair of linked amino acids, there are 6 atoms within the same plane (amide plane)
    • Individual amino acids can be planar on their own individual amide plane
  • Planar peptide bonds can be in Trans or Cis configurations
    • Almost all peptide bonds are in Trans configuration because of steric hindrances in the Cis configuration
    • Amino acid linkages with proline causes steric hindrances in both configurations
  • Bonds in between peptide bonds have angles of rotation
    • phi (φ) is the angle of rotation between nitrogen of amino group and α-carbon
    • psi (ψ) is the angle of rotation between the α-carbon and the carbon atom of carbonyl group
    • Dihedral angle is the measure of rotation about each of the two single bonds – phi or psi
      • usually between -180° and +180°
      • Clockwise rotation is +
    • 3/4 of possible phi and psi combinations are excluded sterically
      • Using Ramachandron diagrams, the precise protein folding can be predicted in large polypeptide chains

Secondary Structure


  • Secondary structure is the special arrangement of amino acid residues along the polypeptide sequence
    • Primary amino acid predicts secondary structure
      • Predictions based on rigidity of peptide bond and restricted set of allowed phi and psi angles
    • Two basic periodic (regularly repeating) secondary structures: α-helix and β-pleated sheets.
    • Secondary structure can be predicted to an extent with different probabilities of a certain amino acid forming part of an α-helix, β-sheet, or reverse turn.

α-Helix

  • α-Helix consist of a tightly coiled backbone with side chains extending outwards
    • Essentially all α-helixes in proteins are right-handed (clockwise) to reduce steric hindrance between –R groups and backbone
      • α-Helix is 1.5 Å wide with 3.6 residues per 360° turn
      • N-H group and C=O hydrogen bonds to stabilize secondary structure with bonds distributed 4 residues apart such that all backbone N-H and C=O groups are bonded
    • α-Helix are very strong and several helixes can super coil with each other to further enhance strength

β-Pleated Sheets

  • β-pleated sheets are more extended and looser in a zigzag structure
    • Adjacent amino acids are stretched 3.5 Å apart
    • Side chains are above and below the peptide bond
    • β-sheets are formed by linking two or more β-strands by hydrogen bonds
      • Can link parallel head to head (with amino terminal ends lined up) or anti-parallel head to tail (with amino terminal lined up with carbonyl terminal)
        • Parallel β-sheets hydrogen bond staggered between amino acids on opposite strand
        • Anti-parallel β-sheets hydrogen bond directly with amino acids on opposite strand
        • Amino acids can have both parallel and anti-parallel structure in the same protein
      • β-sheets can adopt a “flat and twisted” shape

Reverse Turns

  • Reverse turns that compact protein structures are not periodic but rigid
    • Often found on surface of protein and can interact between protein and other molecules
    • Most common structural element is the reverse turn (β-turn, hairpin loop)
      • C=O group of residue i hydrogen bonds with N-H group of residue i+3
    • Omega loops are more elaborate structures responsible for chain reversals

Objectives